The goal of this project is to identify novel inhibitors of the HIV-1 Rev protein for the treatment of AIDS. Rev function is absolutely required for HIV replication. In Phase I, we constructed a reporter plasmid containing the luciferase gene between the HIV-1 exon 6 and exon 7 splice donor and acceptor sites. In this construct, luciferase is expressed only in the presence of Rev. A transient co-transfection assay has been adapted for automated high throughput screening. In Phase II, a fully automated high-throughput screen of 100,000 samples will be carried out, including 50,000 fungal extracts and an equal number of compounds from a medicinal chemistry library. Potential Rev inhibitors will cause a decrease in luciferase activity. Several follow-up assays will be utilized to qualify lead compounds which act specifically on Rev and to exclude cytotoxic compounds. Lead organisms will be refermented, the lead moiety purified and its structure elucidated. The mechanism of action of those entities exhibiting specific Rev-inhibitory properties, e.g. inhibition of Rev-RRE interactions or interaction with cellular factors, will be investigated in vitro. Specific Rev inhibitor lead extracts and compounds will be tested in HIV replication assay systems, representing acute and chronic stages of viral infection. In Phase III, selected leads will be subjected to SAR chemistry and entered into pre-clinical and clinical development. PROPOSED COMMERCIAL APPLICATION: Currently there is no effective therapy for AIDS. Small molecular weight inhibitors of the HIV-1 Rev transactivator, identified as a result of this project, have significant potential as anti-viral agents in the treatment of AIDS.